Process for preparing porous spherical cellulose particles

ABSTRACT

A process for preparing porous, spherical cellulose particles which comprises suspending a cellulose organic ester solution dissolved in a chlorinated hydrocarbon solvent into an aqueous medium to form droplets of the solution, evaporating the chlorinated hydrocarbon solvent in the droplets to form cellulose organic ester spherical particles, and saponifying the cellulose organic ester spherical particles. In the aforesaid process, an acid or alkaki is added and mixed with the cellulose organic ester solution prior to suspending the cellulose organic ester solution into the aqueous medium to control the porosity of the porous, spherical cellulose particles with high reproducibility.

This is a division of application Ser. No. 285,582, filed July 21, 1981,now U.S. Pat. No. 4,390,691, issued June 28, 1983 and the benefits of 35USC 120 are claimed relative to it.

BACKGROUND OF THE INVENTION

(1) Field of the Invention

This invention relates to a process for preparing porous, sphericalcellulose particles.

(2) Description of the Prior Art

Various methods of separating and purifying substances have beendeveloped with the progress in the field of biochemistry. The currentlyused methods as above include ion-exchange chromatography, gelfiltration, affinity chromatography, and the like. Known materials usedin the aforesaid methods include dextran, agarose, polyacrylamide,cellulose, and the like. Many attempts have been made by use of theaforesaid materials so that their properties as a gel filtering materialor ion exchanger may be fully exhibited. A preferable form as the gelfiltering material is a sphere from the standpoints of separationproperty, flow rate and the like.

Few practical applications of cellulose to the gel filtering materialare known at present, but attempts on the preparation of sphericalcellulose particles have been made so far. For example, a process forpreparing cellulose particles from a viscose, or a cuprammoniumsolution, in which cellulose is dissolved, is disclosed in Japanesepatent laid-open specification Nos. 60753/1973, 60754/1973, and5361/1976, and Japanese patent published specification No. 11237/1977.On the other hand, Japanese patent laid-open specification No. 7759/1978discloses a process for preparing spherical cellulose particles fromcellulose acetate.

The process according to Japanese patent laid-open specification No.7759/1978 is so improved as to make it possible to obtain real sphericalcellulose particles having excellent properties as the gel filteringmaterial. The aforesaid process is such that a cellulose triacetatesolution dissolved in a chlorinated hydrocarbon solvent or in a mixedsolvent consisting mainly thereof as a stock solution is added dropwiseinto an aqueous medium with stirring to form suspended particles, andthat the aqueous medium containing the suspended particles is heated toevaporate the solvent and subsequently to form cellulose organic esterspherical particles followed by saponification thereof. The gelfiltering material is often used for the so-called desalting, in which apolymeric substance such as protein is separated from a salt such assodium chloride, urea, ammonium sulfate, or the like. The desalting hasbeen applied on an industrial scale, in which desirable properties asthe gel filtering material require that many samples can be separatedfast with high purity. In order to satisfy the aforesaid properties,some difference is needed between an elution volume of protein and thatof salt. In case that such a gel that has a small difference betweenelution volume of protein and that of salt is used for the treatment ofmuch quantity of sample, the separation ability of the gel isinsufficient and it causes poor purity of eluates, consequently, the gelis hard to be used for industrial scale.

SUMMARY OF THE INVENTION

An object of this invention is to provide an improved process forpreparing porous, spherical cellulose particles usable as a gelfiltering material excellent in separation property due to a sufficientdifference between an elution volume of protein and that of salt byovercoming the aforesaid problems in the process disclosed in Japanesepatent laid-open specification No. 7759/1978.

This invention provides a process for preparing porous, sphericalcellulose particles in which a cellulose organic ester solutiondissolved in a solvent mainly consisting of a chlorinated hydrocarbon issuspended into an aqueous medium to form droplets of the solution, thesolvent in the droplets are evaporated to form cellulose organic esterparticles, and the cellulose organic ester particles are saponified, anacid or alkali being added and mixed with the cellulose organic estersolution prior to suspending the cellulose organic ester solution intothe aqueous medium.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph of Kav values of spherical cellulose particlesobtained by the process of the present invention (E₁, E₂ and E₃), and bythe process of the prior art (CE1) against molecular weight ofpolyethylene glycol used.

FIG. 2 is an elution pattern showing the result of an example ofseparating myoglobin (A) from sodium chloride (B) by use of sphericalcellulose particles obtained in accordance with the process of thepresent invention.

FIG. 3 is an elution pattern showing the result of an example ofseparating cow serum albumin (C) from sodium chloride (D) by use ofspherical cellulose particles obtained in accordance with the process ofthe present invention.

FIG. 4 is an elution pattern showing the result of an example ofseparating myoglobin (A') from sodium chloride (B') by use of sphericalcellulose particles obtained in accordance with the process in the priorart.

FIG. 5 is an elution pattern showing the result of an example ofseparating cow serum albumin (C') from sodium chloride (D') by use ofspherical cellulose particles obtained in accordance with the process inthe prior art.

In FIGS. 2 to 5, OD₂₈₀ represents ultraviolet absorption spectrum(absorbance) at 280 nm.

DETAILED DESCRIPTION OF THE INVENTION AND THE PREFERRED EMBODIMENTS

There are no special limitations to cellulose organic ester used in thepresent invention, and examples thereof may include those obtained byacylation of linter, wood pulp, crystalline cellulose, etc. such ascellulose acetate normally having an acetylation degree of from 45 to62.5%, cellulose acetate butylate normally having an acylation degree offrom 50 to 70%, cellulose butylate normally having a butylation degreeof from 50 to 70.5%, and cellulose acetate propionate normally having anacylation degree of from 48 to 65%.

The solvent usable in the present invention for cellulose organic estermay be selected from those capable of dissolving uniformly the celluloseorganic ester, and examples thereof include chlorinated hydrocarbonsolvents such as dichloromethane, chloroform, carbon tetrachloride,tetrachloroethane, trichloroethane, trichloroethylene,tetrachloroethylene, and mixtures thereof, and mixed solvents of theaforesaid chlorinated hydrocarbon solvents as the major component withother organic solvents such as methanol, ethanol, acetone, andnitromethane.

In the process of the present invention, the acid or alkali may be addedto a solution prepared by dissolving cellulose organic ester in theaforesaid solvent, or to the solvent while cellulose organic ester beingdissolved or prior to dissolution, preferably to the solution preparedby dissolving cellulose organic ester from the standpoints ofreproducibility, and the like.

Examples of acid usable in the present invention include hydrochloricacid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid,phosphoric acid, sulfurous acid, and sulfonic acid such as p-toluenesulfonic acid, preferably inorganic acid.

Examples of alkali usable in the present invention include thehydroxides and alcoholates of alkali metals or alkaline earth metals.

The aforesaid acid or alkali may be used in the form of a solutiondissolved in water, an organic solvent, or a mixed solvent thereof, orin the form of a nonsolvent. The addition amount of the acid or alkaliat its pure state is in the range of from 0.001 to 20% by weight,preferably 0.01 to 15% by weight, more preferably 0.05 to 10% by weightbased on cellulose organic ester.

The concentration of cellulose organic ester in the cellulose organicester solution of the present invention may be selected from the rangeof solubility of cellulose organic ester in the solvent, and ispreferably in the range of from 1 to 15 parts by weight based on 100parts by volume of the solvent, where the relationship between volumeand weight is of l to kg, or ml to g.

In a preferred process of the present invention, acid or alkali is addedto the cellulose organic ester solution and then mixed with agitation ata temperature of from 0° to 45° C., preferably 15° to 40° C. for aperiod of time of from 0.2 to 30 hours, preferably 0.5 to 20 hours, andthe resulting solution is added to an aqueous medium.

The aqueous medium preferably contains a dispersant such as polyvinylalcohol, gelatin, CMC, or the like, or a surfactant of an anionic,cationic, nonionic, ampholytic, or polymer type, or a mixture thereof inorder to maintain the dispersion and shape of droplets therein, and soforth. The amount to be added of the dispersant is suitably in the rangeof from 0.05 to 10% by weight, and the amount to be added of thesurfactant is suitably in the range of from 0.005 to 1% by weight basedon the water in the aqueous medium respectively. An alkali or acid maybe added to the aqueous medium in an amount nearly equivalent to theacid or alkali added to the cellulose organic ester solution.

The cellulose organic ester solution is added to the aqueous medium andsuspended therein to form droplets followed by the evaporation of thesolvent. The evaporation is suitably performed at a temperature higherthan the freezing point of the aqueous medium, preferably 15° C., andless than the boiling point of the chlorinated hydrocarbon solvent.However, the temperature, at which the aforesaid evaporation isperformed, may be raised above the bioling point of the solvent afterthe evaporation of the solvent is performed to such an extent thatcellulose organic ester is solidified.

Cellulose organic ester spherical particles thus obtained are dispersedin, for example, water, alcohol, or a mixture thereof, and aresaponified by adding an aqueous solution hydroxide solution having aconcentration of from 5 to 40% by weight and by stirring at atemperature of from room temperature to 50° C. to form porous, sphericalparticles.

In accordance with the process of the present invention, porous,spherical cellulose particles which have excellent separation propertyin desalting of protein can be obtained. A further feature of thepresent invention is in that the excluded critical molecular weight anddegree of porosity of the porous, spherical cellulose particles can bevaried arbitrarily as desired by varying the amount of the acid oralkali to be added. That is, in accordance with the process of thepresent invention, gel filtering materials having various excludedcritical molecular weights applicable depending on substances to beseparated therewith can also be produced.

Porous, spherical cellulose particles obtained by the process of thepresent invention have such effective pores large in volume and high inmechanical strength as to be usable not only in gel filtration but alsoas a substrate for use in an ion exchanger or affinity chromatography.

The present invention will be described more in detail by the followingexamples, which are not to be construed as limiting its scope. In thefollowing examples, all parts are by weight, and the relationship ofpart by volume to part by weight is equal to that of l to kg or ml to gunless otherwise specified.

The Kav, excluded critical molecular weight, and degree of porosity ofspherical cellulose particles obtained in the following examples aredetermined as follows: Kav: A column having a diameter of 1.6 cm ispacked with spherical cellulose particles swollen with water, which mayhereinafter be referred to as gel, to a height of 50 cm, polyethyleneglycol having a known molecular weight and blue dextran having amolecular weight of 2 millions are added thereto, gel filtration iscarried out at a flow rate of 27 ml/hr by use of water as an elutingsolution, and the elution volumes of each polyethylene glycol and bluedextran are measured by means of a differential refractometer.

Kav is determined by the following equation: ##EQU1## where V_(e) is theelution volume (ml) for eluting polyethylene glycol, V₀ is the hold-upvolume (ml) of the gel and determined as the elution volume (ml) foreluting blue dextran; and V_(t) is the total bed volume (ml), anddetermined as the product of the cross sectional area of the column bythe height of the gel bed. Excluded critical molecular weight:

Excluded critical molecular weight is determined as the molecular weightof polyethylene glycol at a turning point of a curve obtained byplotting Kav values of the gel as an object obtained by use of variouspolyethylene glycols having known molecular weights respectively againstrespective molecular weights of the polyethylene glycols. Degree ofporosity:

A degree of porosity as a measure of porosity is determined according tothe following equation: ##EQU2##

It can be said that the greater the degree of porosity, the higher theporosity.

EXAMPLE 1

160 parts of cellulose acetate having an acetylation degree of from 60to 61% (marketed by Celanese Corp. under the trademark of CelluloseTriacetate KB 175) are dissolved in 2,000 parts by volume ofdichloromethane, and 2 parts by volume of 36 weight % hydrochloric acidis added thereto at 15° C. to be stirred for 10 hours. Thereafter, theresulting mixture is added to an aqueous medium consisting of 2,700parts of water, 27 parts of polyvinyl alcohol, and 2.7 parts of sodiumdodecylbenzenesulfonate at 35° C. After stirring further for some periodof time, nitrogen gas is passed into the gaseous phase over the liquidsurface to evaporate dichloromethane.

After the completion of evaporation of the total amount ofdichloromethane gas, cellulose acetate particles thus formed arefiltered and washed with water. Wet particles sucked thoroughly with asuction funnel are added to 1600 parts by volume of 75 volume %water-containing methanol, suspended, stirred at 50° C. for 30 minutes,and cooled to a temperature lower than 30° C. To the resulting mixture,398 parts of 20 weight % aqueous sodium hydroxide solution are added,and the mixture is stirred at a temperature of from 20° to 30° C. for awhole day for saponification. The resulting mixture is neutralized withacetic acid and filtered to obtain spherical cellulose particles, whichare washed thoroughly with water and classified to a particle size offrom 50 to 100 microns in diameter to obtain a final product.

The product thus obtained is subjected to measurement of Kav valuesthereof for respective molecular weights of various polyethyleneglycols, and the Kav values thus obtained are plotted against respectivemolecular weights to obtain a curve E₁ as shown in FIG. 1. The curve E₁shows that the excluded critical molecular weight of the product isabout 2,000. The curve E₁ also shows a great difference between Kavvalues, that is elution volumes of a compound having a molecular weightgreater than 2,000 and one having a molecular weight of 1,000. Thisshows high separation properties between two compounds as above, thatis, excellent performance of desalting for separating protein from asalt of a low molecular weight.

The degree of porosity for the product was 0.47.

EXAMPLE 2

Separation of myoglobin having a molecular weight of 17,000 from sodiumchloride having a molecular weight of 58 by use of the product inExample 1 is carried out in the following manner to determine R, degreeof separation. A column having an inner diameter of 1.5 cm is packedwith a gel (the product obtained in Example 1) to a height of 49 cm. Asolution prepared by dissolving 50 mg of myoglobin and 250 mg of sodiumchloride in 2 ml of 0.05M aqueous ammonium formate solution is added tothe packed column for carrying out gel filtration by use of 0.05Maqueous ammonium formate as an eluting solution at a flow rate of 28ml/hr. The eluting solution is dispensed by every 10 ml by a fractioncollector. Myoglobin is determined by ultraviolet absorption spectrum at28 nm, and sodium chloride is determined by determination of Cl⁻ ion bymeans of silver nitrate titrimetric determination. Amounts of solutesfor respective fractions are plotted against elution volume as shown inFIG. 2, in which A represents myoglobin, and B represents sodiumchloride. From the graph in FIG. 2, R, degree of separation isdetermined by the following equation: ##EQU3## where V₁ and V₂ areelution volume of a solute 1 (sodium chloride) and a solute 2(myoglobin) respectively, and W₁ and W₂ are respective widthes on thebase line of the peaks of the solutes 1 and 2.

The greater R is, the higher the separating property. R thus obtained isequal to 3.6.

EXAMPLE 3

Separation of cow serum albumin is an aqueous solution from sodiumchloride by use of the product in Example 1 is carried out in thefollowing manner. A column having an inner diameter of 2.6 cm is packedwith a gel filtering material (the product obtained in Example 1) to aheight of 55.5 cm. 80 ml of an aqueous solution having an albuminconcentration of 5% (weight/volume) and a sodium chloride concentrationof 15% are added to the packed column for carrying out gel filterationby use of water as an eluting solution at a flow rate of 95 ml/hr. Theelution solution is dispensed by every 10 ml by a fraction collector.Albumin is determined by ultraviolet absorption spectrum at 280 nm, andsodium chloride is determined by determination of Cl⁻ ion by means ofsilver nitrate titrimetric determination. Amounts of solutes forrespective fractions are plotted against elution volume as shown in FIG.3, in which C represents albumin, and D represents sodium chloride.Taking into considerations the graph in FIG. 3, the fractions of from11th to 20th are collected to be 100 ml in total. The eluting solutionthus collected has an albumin concentration of 3.9% and a sodiumchloride concentration of 0.14%. This shows that recovery percentage foralbumin is 98% and the concentration of sodium chloride is decreasedfrom 15% to 0.14% with excellent desalting performance.

COMPARATIVE EXAMPLE

Procedures of Example 1 are repeated except that no hydrochloric acid isadded to a dichloromethane solution of cellulose acetate to obtainspherical cellulose particles, properties of which are described asfollows.

Relationship of Kav value with the molecular weight of polyethyleneglycol is shown by a curve CE1 in FIG. 1, resulting an excluded criticalmolecular weight of 1000, and in a degree of porosity of 0.29.

Comparison of the above results with the results in Example 1 apparentlyshows that the product in Example 1 has a greater excluded criticalmolecular weight, a higher degree of porosity, and a higher separatingproperty as shown by comparison of Kav at a molecular weight greaterthan 2,000 with that at a molecular weight of from 200 to 300 betweenthe curves E1 and CE1 respectively.

The product obtained in Comparative Example 1 is subjected to separationof myoglobin from sodium choride in the same manner as in Example 2. Theresult thus obtained is shown in FIG. 4, in which A' representsmyoglobin, and B' represents sodium chloride, R being equal to 2.0.Comparison of the above value of R with that (3.6) or R in Example 3shows that the product of the present invention has higher separatingproperty.

Separately, the product obtained in Comparative Example 1 is subjectedto separation of cow serum albumin from sodium chloride in the samemanner as in Example 3. The result thus obtained is shown in FIG. 5, inwhich C' represents albumin, and D' represents sodium chloride. Thegraph in FIG. 5 shows a poor separation of albumin from sodium chloride.The fractions of from 11th to 20th are collected to show an albuminrecovery percentage of 98% and a sodium chloride concentration of 5%.Comparison of the result thus obtained with that in Example 3 shows thatthe product of the present invention has much higher desalting propertyand separating property.

EXAMPLE 4

The excluded critical molecular weight and degree of porosity for gelparticles (porous, spherical cellulose particles) prepared in the samemanner as in Example 1 with varied amounts of hydrochloric acid to beadded to the dichloromethane solution of cellulose acetate are shown inTable 1.

                  TABLE 1                                                         ______________________________________                                        Sample No.     1        2       3      4                                      ______________________________________                                        Amount of      5        2.5     1      0.5                                    hydrochloric acid                                                             Excluded critical                                                                            3,000    2,500   1,200  1,000                                  molecular weight                                                              Degree of porosity                                                                           0.56     0.50    0.45   0.40                                   ______________________________________                                    

Table 1 apparently shows that both excluded critical molecular weightand degree of porosity can arbitrarily be varied by varying the amountto be added of hydrochloric acid.

Relationship of Kav with molecular weight for gel particles of thesample No. 1 in Table 1 is shown by a curve E₂ in FIG. 1. Comparison ofcurve E₂ with curve CE₁ apparently shows that gel particles of sampleNo. 1 in Example 4 have higher separating property.

EXAMPLE 5

Such properties as excluded critical molecular weight and degree ofporosity of cellulose particles prepared in the same manner as inExample 1 except for the use of varied amounts of sulfuric acid areshown in Table 2.

                  TABLE 2                                                         ______________________________________                                        Sample No. 1          2           3                                           ______________________________________                                        Amount of  0.5 part by                                                                              0.5 part by 0.5 part by                                 sulfuric acid                                                                            volume (36%                                                                              volume (12% volume (4%                                             by weight) by weight)  by weight)                                  Excluded critical                                                                        1,500      1,200       1,000                                       molecular weight                                                              Degree of  0.40       0.37        0.33                                        porosity                                                                      ______________________________________                                    

Relationship of Kav with molecular weight for gel particles of thesample No. 1 in Table 2 is shown by a curve E₃ in FIG. 1.

Table 2 apparently shows that both excluded critical molecular weightand degree of porosity can arbitrarily by varied by varying the amountto be added of sulfuric acid, and that gel particles of the sample No. 1in Example 5 have higher separating property as shown by comparison ofcurve E₃ with curve CE₁.

EXAMPLE 6

Procedures of Example 1 are repeated except that 2 parts by volume of 40weight % aqueous sodium hydroxide solution is added to the celluloseacetate solution instead of adding hydrochloric acid to thedichloromethane solution of cellulose acetate, and stirred at 30° C. for20 hours prior to addition thereof to the aqueous medium to obtaincellulose particles, the excluded critical molecular weight and degreeof porosity of which are 2,000 and 0.38 respectively. Relationship ofKav with molecular weight is almost the same as that of curve E₁ in FIG.1.

EXAMPLE 7

Procedures of Example 6 are repeated except for the use of 2 parts byvolume of 20 weight % aqueous sodium hydroxide solution to obtaincellulose particles, the excluded critical molecular weight and degreeof porosity of which are 1,500 and 0.36 respectively.

EXAMPLE 8

100 parts of cellulose acetate having an acetylation degree of 55% aredissolved in a mixed solvent of 700 parts by volume of dichloromethaneand 300 parts by volume of acetone, and one part by volume of 35 weight% hydrochloric acid is added thereto at 35° C. to be stirred for 10hours. Thereafter, the resulting mixture is added to an aqueous mediumconsisting of 3,000 parts of water and 150 parts of gelatin at 35° C.After stirring further for some period of time, nitrogen gas is passedinto the gaseous phase over the liquid surface. Dichloromethane isremoved at 35° C., and the temperature of the solution is raised up to55° C. to remove acetone. Particles thus formed are filtered and washedwith water. The resulting particles are sucked thoroughly with a suctionfunnel, added to 1,000 parts by volume of 75 volume % water-containingmethanol to be suspended therein, stirred at 50° C. for 30 minutes,cooled to a temperature lower than 30° C. To the resulting mixture, 250parts of 20 weight % aqueous sodium hydroxide solution are added, andstirred at a temperature of from 20° to 30° C. for a whole day forsaponification. The resulting mixture is neutralized with acetic acidand filtered to obtain spherical cellulose particles, which are washedthoroughly with water and classified to a particle size of from 50 to100 microns in diameter to obtain a final product. The product thusobtained has such properties that the excluded critical molecular weightis about 2,000, relationship of Kav with molecular weight is almost thesame as that of curve E₁ in FIG. 1, and that degree of porosity is 0.47.

EXAMPLE 9

160 parts of cellulose acetate butylate having 13% of acetyl group and37% of butyl group are dissolved in a mixed solvent of 1,400 parts byvolume of dichloromethane and 600 parts by volume of methyl ethylketone, and 2 parts by volume of 36 weight % hydrochloric acid are addedthereto at 15° C. to be stirred for 10 hours. Thereafter, the resultingmixture is added to an aqueous medium consisting of 2,700 parts ofwater, 27 parts of polyvinyl alcohol, and 2.7 parts ofdodecylbenzenesulfonic acid at 35° C. After stirring further for someperiod of time, nitrogen gas is passed into the gaseous phase over theliquid surface to evaporate dichloromethane. The temperature of thesolution is then raised up to 80° C. to remove methyl ethyl ketone andto obtain spherical particles of cellulose acetate butylate. Thereafter,procedures of Example 1 are repeated to obtain spherical celluloseparticles. The cellulose particles thus obtained have such propertiesthat excluded critical molecular weight is 2,000, relationship of Kavwith molecular weight is almost the same as that of curve E₁ in FIG. 1,and that degree of porosity is 0.47.

What is claimed is:
 1. Porous, spherical cellulose particles with highreproducibility, produced by dissolving a cellulose organic ester in asolvent consisting essentially of a chlorinated hydrocarbon, adding0.001-20 wt.% (based on said cellulose organic ester) of an acid oralkali to the dissolved cellulose organic ester solution, mixing theresultant solution at a temperature from 0° to 45° C. for a period oftime from 0.2 to 30 hours, suspending said cellulose organic estersolution in an aqueous medium to thereby form droplets, removing saidsolvent by evaporation to form cellulose organic ester particles, andsaponifying said cellulose particles, the excluded critical molecularweight and the degree of porosity of the porous, spherical celluloseparticles being capable of being varied as desired by varying the amountof the acid or alkali to be added.
 2. Particles according to claim 1wherein the concentration of said cellulose organic ester in thesolution of cellulose organic ester dissolved in said solvent is in therange of from 1 to 15 parts by weight based on 100 parts by volume ofthe solvent.
 3. Particles according to claim 1 wherein said solventconsists essentially of at least one chlorinated hydrocarbon selectedfrom dichloromethane, trichloromethane, carbon tetrachloride,trichloroethane, tetrachloroethane, trichloroethylene andtetrachloroethylene.
 4. Particles according to claim 1 wherein saidsolvent is a mixed solvent containing chlorinated hydrocarbon as themajor component and at least one material selected from methanol,ethanol, acetone and nitromethane.
 5. Particles according to claim 1wherein an acid is added.
 6. Particles according to claim 5 wherein saidacid is an inorganic acid.
 7. Particles according to claim 5 whereinsaid acid is at least one selected from hydrochloric acid, hydrobromicacid, hydrofluoric acid, sulfuric acid, sulfurous acid, phosphoric acidand nitric acid.
 8. Particles according to claim 5 wherein said acid issulfonic acid.
 9. Particles according to claim 1 wherein an alkali isadded.
 10. Particles according to claim 9 wherein said alkali is analkali metal hydroxide or an alkaline earth metal hydroxide. 11.Particles according to claim 1 wherein said acid or alkali is added toan amount of from 0.001 to 20% by weight based on said cellulose organicester.
 12. Particles according to claim 11, wherein said acid or alkaliis added in an amount of from 0.05 to 10% by weight based on saidcellulose organic ester.
 13. Particles according to claim 1 wherein saidacid or alkali is added and mixed with the cellulose organic estersolution after the cellulose organic ester is dissolved in saidchlorinated hydrocarbon solvent.
 14. Particles according to claim 12wherein after dissolving said acid or alkali in the cellulose organicester solution, the resulting solution is mixed with stirring at atemperature of from 0° to 45° C. for a period of time of from 0.2 to 30hours, and added to the aqueous medium to be suspended therein. 15.Particles according to claim 1 wherein said acid or alkali is added andmixed with the solvent before the cellulose organic ester is dissolvedin said chlorinated hydrocarbon solvent.
 16. Porous, spherical celluloseparticles with high reproducibility, produced by dissolving a celluloseorganic ester in a solvent consisting essentially of a chlorinatedhydrocarbon, mixing the resultant solution at a temperature from 0° to45° C. for a period of time of from 0.2 to 30 hours, suspending saidcellulose organic ester solution in an aqueous medium to thereby formdroplets, removing said solvent by evaporation to form cellulose organicester particles, and saponifying said cellulose particles, 0.001 to 20wt% (based on said cellulose organic ester) of an acid or alkali beingadded to said solvent while the cellulose organic ester is beingdissolved or prior to said dissolution, the excluded critical molecularweight and the degree of porosity of the porous, spherical celluloseparticles being capable of being varied as desired by varying the amountof the acid or alkali to be added.